Immunoelectrophoresis gel tray

ABSTRACT

SUBSEQUENT DIFFUSION OF THE ANTI-SERUM TO PRECIPITATE PROTEINS.   AN ELECTROPHORESIS TRAY HAVING WIRE-LIKE ELEMENTS DISPOSED THEREIN FOR INJECTING INTO A GEL CARRIED BY THE TRAY, SERUM AND ANTI-SERUM TO PROVIDE FOR ELECTROPHORESIS AND

y 1973 s. D. RAINS 3,736,100

IMMUNOELECTROPHORESIS GEL TRAY Filed May 24. 1971 STEPHEN D. RAINSINVENTOR.

BERNARD D. BOGDON ATTORNEY United States Patent O 3,736,100INIMUNOELECTROPHORESIS GEL TRAY Stephen D. Rains, Henrietta, N.Y.,assignor to Bausch & Lomb Incorporated, Rochester, NY.

Filed May 24, 1971, Ser. No. 146,388 Int. Cl. B01k /00; G01n 31/02,33/16 U.S. Cl. 23-253 R 11 Claims ABSTRACT OF THE DISCLOSURE Anelectrophoresis tray having wire-like elements disposed therein forinjecting into a gel carried by the tray, serum and anti-serum toprovide for electrophoresis and subsequent diifusion of the anti-serumto precipitate proteins.

BACKGROUND OF THE INVENTION (1) Cross-references to related applicationThis invention is related to the invention described in the concurrentlyfiled and copending applications entitled (l) Method of and Apparatusfor Depositing a Fluid in a Gel, 'Ser. No. 146,262 for inventors StephenD. Rains and Leon L. Wheeless, Jr. and (2) -EIectr0- phoresis System andGel Frame, Ser. No. 146,387 for inventor Stephen D. Rains.

(2) Field of the invention This invention relates to an apparatus forcarrying gel for immunoelectrophoresis analysis of serums and antiserumsand more particularly to a self-contained unit for supporting the gelincluding apparatus for individually introducing serum and anti-serum.

(3) Description of the prior art The practice of immunolectrophoresishas been known for many years and certain difliculties have beenexperienced in the practice, with particular regard to introducingserums and anti-serums into a gel in order to carry out the process.Frequently, contamination of the serums and anti-serums between eachother has resulted while attempting to introduce the two into the mediumfor carrying out the electrophoresis processes and subsequentprecipitation processes. These difficulties have lead to results whichare undependable, costly and time-consuming. Quite often, it isnecessary to repeat the process in order to assure that accurate resultsare obtained.

SUMMARY OF THE INVENTION This invention provides an apparatus forexpeditiously and inexpensively carrying out the process of immunmelectrophoresis with dependable accurate results. In the hereinbeforementioned patent application entitled Method of and Apparatus forDepositing a Fluid in a Gel for Rains et al., there is described, indetail, a method for introducing fluids into a gel for analyzingchemical constituents. Further, R. I. Wieme in Agar Gel Electrophoresis;published by Elsevier Publishing Company in 1965, describes, in detail,the particular processes of electrophoresis as well asimmunoelectrophoresis. The embodiments disclosed therein generally showa wire-like element which extends across the frame for introducing afluid into a gel contained within the frame. It is described in thespecification that the wirelike element may be constructed from variousmaterials which will produce a wire-like element that may be anythingfrom rigid to flexible and, also, anything therebetween. The onlydefinite requirement is that the wirelike element should be able to bepulled from the gel and generally remain within the path that itinitially defined within the gel. Accordingly, in the present de-3,736,100 Patented May 29, 1973 lCC BRIEF DESCRIPTION OF THE DRAWINGSFIG. 1 is a perspective view of an apparatus according to the principlesof the present invention;

FIG. 2 is an enlarged cross-sectional view of the apparatus of FIG. 1taken along the line 22 of FIG. 1;

FIG. 3 is a visual pattern of the precipitated proteins after completionof the process of immunoelectrophoresis; and

FIG. 4 is a cross-sectional view of another embodiment of the apparatusaccording to the principles of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In FIG. 1 there isdisclosed a gel frame 10 having a recessed area 12 defined by parallellateral walls 14 and 16 and transversely disposed walls 18 and 20 forcarrying any gel suitable for use in the process of electrophoresis.Extending upward from the bottom portion of the recessed area 12 are aplurality of protrusions 22, 22 and 2 of similar construction. Theprotrusions 22, 22' and 22" are ideally formed as an integral part ofthe gel frame 10 and are generally rectangular in shape. Disposed atopposite sides of each of the protrusions 22, 22 and 22" are respectiveapertures 24 and 26-, 24 and 26 and 24 and 26". The series of apertures24, 26, 24', 26, 24" and 26" extend through the gel frame 10 from therecessed area '12 to the underside 28 of the gel frame 10.

As best seen in FIG. 2, inverted U-shaped' wire-like elements 30, 30'and 30" extend respectively through the series of apertures 24, 26, 24,26 and 24", 26" to bridge the respective series of protrusions 22, 22and 22!! In its use the gel frame =10 is turned to its underside 28 sothat an operator would be able to dispose a serum droplet onto one endof each of the U-shaped wire-like elements 30, 30' and 30" andthereafter extract the Wire-like elements from the gel frame 10 bypulling on the end of the U-shaped wire-like element opposite thedroplet. This action will cause the droplet to follow and, consequently,fill the void left in the gel by the extraction of the U-shapedwire-like elements 30, 30' and 30" from the gel retained in the recessedarea 12. Once the serum is introduced into the gel the frame iselectrically connected by any suitable means and the process ofelectrophoresis is conducted. After the serum migration has transpiredacross the top surface of the gel frame in the direction parallel to thewalls 14 and 16, the gel frame is ready for the further process and theoperation of immunoelectrophoresis.

To this end the gel frame 10 has included therein aligned recesses 32and 34 extending from the transverse walls 1 8 and 20 outward to theouter periphery of the gel frame 10'. Additionally, included in the pathof the recess 34 is a reservoir 36 having any suitable shape andextending, for example, to the same depth as the recessed area 12. Itwill be appreciated that a plurality of recesses 32 and 34 and reservoir36 may be incorporated as shown by recesses 32' and 34' and reservoir36'. Extending through the gel carried in the recessed area 12 andaligned to the recesses 32 and 34 is a wire-like element 38. Likewise, aWire-like element 38' extends through recesses 32 and 34'. The Wirelikeelement 38 is best made of an electrically non-conductive material,since it is in place in the gel while the electrical power is appliedfor the electrophoresis process.

After the hereinbefore mentioned process for electrophoresis of a serumhas been conducted, an anti-serum of known composition to conductparticularly desired tests is disposed in the respective reservoirs 36and 36' and the wire-like elements 38 and 38' are extracted.Correspondingly, upon extraction of the wire-like elements 38 and 38,the anti-serum fills the void left by the respective wire-like elementsas described in the hereinbefore mentioned Rains et al. patentapplication entitled Method of and Apparatus for Depositing a Fluid in aGel.

After the above-mentioned steps have been completed it is only necessaryto incubate the gel in an evironment suitable for diffusion of serum andanti-serum, and for precipitation of the antigen-antibody complexes.After the prescribed amount of time has elapsed the serum and antiserumwill have reacted to each other and an analysis can quickly be made bysimple visual interpretation. The results of the reaction of the serumto the anti-serum can readily be seen in FIG. 3. The precipitin arcsthat appear take on an appearance of a series of overlapping crescentsdesignated as numeral 40 in the drawing of FIG. 3.

A further embodiment of the present invention is disclosed in FIG. 4.Specifically, a gel frame 42 is shown having a recessed portion 44therein. A plurality of passageways 46, 48 and 50 are located in the gelframe 42 so as to communicate the lower surface 52 of the gel frame withthe bottom 54 of the recess 44. A plurality of wirelike elements '56, 8and 60 are positioned in the passageways 46, 48 and 50, respectively, sothat they do not protrude above the upper surface 62 of the gel frame42. A gel 64 is then introduced into the recessed portion 44 of the gelframe 42 and allowed to set. Once the gel 64 has set a portion of theresidual gel that remains in the conical portions of the passageways 46,48 and 50, respectively, may be removed therefrom. The removal of thesmall amount of residual gel is necessary to allow a fluid sample to bedeposited in the conical shaped sections of the passageways 46, 48 and50. The fluid sample may be deposited by any known acceptable mannersuch as, for instance, was disclosed in the concurrently-filed andcopending application Method of and Apparatus for De positing a Fluid ina Gel for inventors Stephen D. Rains and Leon L. Wheeless, Jr. After thefluid sample has been deposited in the passageways the wire-likeelements may be withdrawn therefrom. The fluid, as previously described,is caused to flow into the voids left by the extraction of the wire-likeelements. The hereinbefore mentioned patent application entitled Methodof and Apparatus for Depositing a Fluid in a Gel specifically describesthis phenomenon.

It will be appreciated that although specific articles have been givenspecific identifications there are a great number of other variations ofthese articles that would work equally as well. For instance, the meansfor introducing the fluid into the gel has been called a wire-likeelement, although anything that resembles a wire, such as a strand ofhair, a nylon or plastic filament, or even a string, would work equallyas well. Further, the wire-like element may be either rigid or flexibleand it is not necessarily limited to a cross-section of circular shape.A strip of material such as paper or plastic, for example, can alsoserve as the wire-like element, as well as can probes of triangular,square or any of the cross-sectional shape.

I claim:

1. An apparatus for use in the process of immunoelectrophoresis wheregel is useable as a support medium, comprising:

' a frame defining a recessed portion therein for carrying gel, saidrecessed portion having at least one aperture communicating the recessedportion with an outer surface of the frame;

at least one first filament, said first filament extending through theaperture to define a first path for fluid through the aperture and gelcarried within the recessed portion to facilitate the introduction ofchemical constituents into gel carried within the recessed portion; and

at least one second filament, said second filament disposed within andtraversing at least part of the length of the recessed portion to definea second separate path for fluid to facilitate the introduction ofadditional chemical constituents into gel carried within the recessedportion.

2. The apparatus as defined in claim 1, wherein the recessed portion hastwo apertures, and the first filament is U-shaped and extends throughsaid two apertures in the recessed portion of the frame to define aU-shaped first path for fluid.

3. The apparatus as defined in claim 2, further comprising support meansfor supporting the U-shaped first filament within the recessed portion.

4. The apparatus as defined in claim 3, wherein the support means forsupporting the U-shaped first filament is a protruding member disposedbetween the apertures in the recessed portion.

5. The apparatus as defined in claim 4 wherein the protruding member isintegral with the frame.

6. An apparatus for use in the process of immunoelectrophoresis where agel is useable as a support medium, comprising:

a frame defining a recessed portion therein to carry gel, said recessedportion having a plurality of apertures communicating the recessedportion with an outer surface of the frame;

at least one U-shaped element, said element extending through twoapertures to define a first continuous path for fluid through the twoapertures and gel carried within the recessed portion;

at least one support member for said U-shaped element, said supportmember disposed between said U-shaped element and the recessed portionto provide support for said U-shaped element; and I at least oneelongated straight element positioned adjacent said U-shaped element andwithin the recessed portion to define a second continuous path for fluidwithin the gel separate from the first continuous path.

7. An apparatus for use in the process of immunoelectrophoresis where agel is useable as a support medium, comprising:

a frame defining a recessed portion therein for carrying gel, saidrecessed portion having at least one aperture communicating the recessedportion with an outer surface of the frame;

at least one first substantially straight element extending through saidaperture to define a first path for fluid through the aperture and gelcarried within the recessed portion to facilitate the introduction ofchemical constituents into the gel carried within the recessed portionat the first path; and

at least one second substantially straight element disposed within andtraversing at least part of the length of the recessed portion to definea second separate path for fluid to facilitate the introduction ofadditional chemical constituents into gel carried within the recessedportion at the second path.

8. An apparatus for use in the process of immunoelectrophoresis wheregel is carried as a support medium for analysis, comprises:

a frame defining a recessed portion therein for carrying a gel, saidrecessed portion having at least one aperture communicating the recessedportion with an outer surface of the frame; a gel carried within the'recessed portion of the frame; at least one first filament, said firstfilament disposed in the gel and extending through the aperture todefine a first path for fluid through the aperture and gel carriedwithin the recessed portion to facilitate the introduction of chemicalconstituents into the gel carried within the recessed portion; and atleast one second filament, said second filament disposed within the geland traversing at least part of the length of the recessed portion todefine a second separate path for fluid to facilitate the introductionof additional chemical constituents into the gel carried within therecessed portion. 9. The apparatus as defined in claim 8, wherein thesecond filament is elongated and substantially straight.

10. The apparatus as defined in claim 8, wherein the recessed portionhas two apertures and the first filament References Cited UNITED STATESPATENTS 4/1968 Saravis et a1. 23-230 B X 6/1968 Saravis 23-230 B MORRISO. WOLK, Primary Examiner R. M. REESE, Assistant Examiner US. Cl. X.R.

23-230 B; 204-180 G, 299 R

